Details zur Publikation |
| Kategorie | Textpublikation |
| Referenztyp | Zeitschriften |
| DOI | 10.1016/j.jprot.2008.11.018 |
| Titel (primär) | Standardized peptidome profiling of human cerebrospinal fluid by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry |
| Autor | Bruegel, M.; Planert, M.; Baumann, S.; Focke, A.; Bergh, F.T.; Leichtle, A.; Ceglarek, U.; Thiery, J.; Fiedler, G.M. |
| Quelle | Journal of Proteomics |
| Erscheinungsjahr | 2009 |
| Department | METABOX; PROTEOM |
| Band/Volume | 72 |
| Heft | 4 |
| Seite von | 608 |
| Seite bis | 615 |
| Sprache | englisch |
| Keywords | Cerebrospinal fluid; MALDI-TOF; Mass spectrometry; Peptidome; Proteome; Standardization |
| Abstract | Peptidome profiling of human cerebrospinal fluid (CSF) is a promising tool to identify novel disease-associated biomarkers. Our aim was to develop a standardized protocol for reproducible peptidome profiling of CSF using magnetic bead (MB) separation followed by MALDI-TOF MS. Peptidome fractionation and profiling of CSF were performed using MBs with different surface functionalities. We investigated exogenous variables (storage conditions, freeze-thaw-cycles) and endogenous interferences (albumin, immunoglobulin, blood, leukocytes) in pooled CSF samples. We detected approximately 500 signals with an S/N ratio > 10 and an overlap frequency of about 40% in non-pathological CSF. Within- and between-day imprecisions in relative signal intensities ranged from 3 to 28% and 7 to 47%, respectively. CSF storage at room temperature for up to 6 h and at 4 °C for up to 3 days did not significantly influence the mass spectra. Consecutive freeze-thaw-cycles significantly affected the mass spectra. High albumin and immunoglobulin content altered the CSF preparation using MB-HIC C8 beads. Blood contamination showed no effect on mass spectra up to a hemoglobin concentration of 0.075 µmol/L. The presence of leukocytes up to a cell number of 30 Mpt/L did not affect mass spectra. Our reliable pretreatment protocol allows standardization of preanalytical modalities and thereby enables reproducible peptidome profiling of human CSF using MB separation followed by MALDI-TOF MS. |
| dauerhafte UFZ-Verlinkung | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=9838 |
| Bruegel, M., Planert, M., Baumann, S., Focke, A., Bergh, F.T., Leichtle, A., Ceglarek, U., Thiery, J., Fiedler, G.M. (2009): Standardized peptidome profiling of human cerebrospinal fluid by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry J. Proteomics 72 (4), 608 - 615 10.1016/j.jprot.2008.11.018 |
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